Anti-ISWI (acetyl K753)抗体(ab10748)

概述

The specificity of the antibody can be summarized as follows:
1. On Western blots probing recombinant ISWI which had been (or not)
acetylated at K753 by GCN5, the antibody recognizes the acetylated ISWI
with very high selectivity. No reaction with unacetylated ISWI has been observed.
2. Unexpectedly, there is a significant cross-reactivity with GCN5 under
acetylation conditions (we assume autoacetylated GCN5). As shown in the figure, this cross-reactivity is low, since conditions can be found easily where the GCN5 band on Western blots is not observed.
3. In whole cell extracts the antibody cross-reacts with a number of small molecular weight bands. A faint signal at the position of ISWI is detected as well, which we think corresponds to ISWI since it is not present if ISWI is ablated using RNAi. The weakness of the signal may simply indicate that there is very little epitope present in these cells.
In summary, we are confident that the antibody can be used on purified ISWI (e.g.
ISWI immunoprecipitated from a cruder source).

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应用

Fast track antibodies constitute a diverse group of products that have been released to accelerate your research, but are not yet fully characterized. They have all been affinity purified and show high titre values against the immunizing peptide (by ELISA).
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应用

Ab评论

说明

ELISA

Use at an assay dependent dilution. This antibody gave a positive result in ELISA against the immunizing peptide (ab16064). It gave a negative result in ELISA against the non-modified equivalent peptide (ab96245). This indicates that it is specific for the modified peptide.

ICC/IF

Use at an assay dependent concentration. PubMed: 22949616

靶标

相关性

ISWI is a component of the nucleosome remodeling factor complex (NURF), a protein complex that facilitates the perturbation of chromatin structure in vitro in an ATP-dependent manner.
The hydrolysis of ATP during the remodeling of chromatin is likely to be mediated by ISWI, releasing inorganic phosphate. It is also a component of the ATP-utilizing chromatin assembly and remodeling factor (ACF) and of the chromatin accessibility complex (CHRAC). This subunit may serve as the energy-transducing component of chromatin-remodeling machines.

Lane 1 : As aboveLane 2 : Recombinant ISWI (200 ng) was acetylated using 125ng of hGCN5 and a final concentration of 6 µM of 3H-AcCoA.Lane 3 : Recombinant ISWI (200 ng) was acetylated using 125ng of hGCN5 and a final concentration of 6 µM of 3H-AcCoA.Lane 4 : As above

Recombinant ISWI (200 ng) was acetylated using 125 ng (lanes 2 and 3) of hGCN5 and a final concentration of 6 µM of 3H-AcCoA.

Acetylated proteins were detected by western blotting using the acetylated ISWI (K753) polyclonal antibody (ab10748), diluted 1/2500 (lanes1 and 2) or 1/500 (lanes 3 and 4) in 3% milk 0.1% T-PBS. In lanes 3 and 4, 1/5th of the reaction was loaded.

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Have you had the opportunity to test the antibody in Western blot as well? Indeed, as you have a positive control, I can recommend strongly to test the antibody in Western blot to see whether the antibody has retain...

Thank you for your enquiry. I am pleased that you have expressed an interest in this product. This is an in house product and the images are derived from a collaborator. We do not as yet possess any specific published references relating to this produc...